International Journal of

Applied Sciences & Engineering

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RESEARCH ARTICLE

Effect of Traditional Smoking and Cleaning of Milk Equipment with Different Plants and Herbs
on Keeping Quality and Organoleptic Properties of Raw Milk in Ethiopia
Zerihun Asefa1 and Esayas Abrha2
1,2
Ethiopian Institute of Agricultural Research, Holeta Agricultural Research Center, PO box 2003, Addis Ababa, Ethiopia

AR T IC L E I NFO AB S T RA C T

Received: December 12, 2021 The high moisture content, micronutrient content, and pH near to neutral make
Revised: January 21, 2022 cow milk an ideal medium for the growth of spoilage as well as pathogenic
Accepted: February 10, 2022 microorganisms. This study aims to evaluate the traditional practices of milk
utensils cleaning and smoking with different herbs and plants to improve the
Keywords:
keeping quality as well as organoleptic properties of raw milk in rural parts and
Smoking
the agro-pastoral community of Ethiopia. In this study three, the most common
Organoleptic
cleaning herbs and three smoking plants traditionally practiced in the
Keeping quality
community were used in combinations. Accordingly, nine treatments and one
Cleaning
control milk utensil cleaned with only water were used. Raw milk was stored in
each utensil and microbial growth, physicochemical properties of pH and
treatable acidity, and organoleptic properties of raw milk were evaluated at
different times of storage at room temperature. The change in pH and treatable
acidity of raw milk in T1, T4, and T7 were no significant changes between 0,
12, and 24 hrs of storage. The microbial growth of total plate count, total
coliform, and lactic acid bacteria were no significant change during 0, 12, and
24 hrs of storage in treatment T1, T4, and T7. The organoleptic property of odor,
test, and overall acceptability was shown a significant difference between
treatments. Utilization of treatments mentioned as T1, T4, and T7 was better
*Corresponding Address: keeping quality, as well as organoleptic property on raw milk during storage at
zafuase@gmail.com, room temperature. So, they are better for the rural community of Ethiopia to
+251911552657 keep milk safe and healthy until delivered to the market.

Cite This Article as Asefa Z, Abrha E, 2021. Effect of traditional smoking and cleaning of milk equipment with
different plants and herbs on keeping quality, and organoleptic properties of raw milk in Ethiopia. Inter J Appl Sci Engr,
9(1): 26-30. www.ijapscengr.com

INTRODUCTION mostly transferred from generation to generation through

Milk is a perishable commodity that is spoiled easily
and quickly. High moisture content, micronutrient content,
and pH nears to neutral make it an ideal medium for the
growth of spoilage and pathogenic bacteria and poses
serious human health threats (Wanjala et al., 2016b).
Smoking of milking equipment with herbs is used in
pastoral communities of Kenya to disinfect milking
equipment, to improve the flavor of milk, and to extend
their shelf life (Wafula et al., 2016). In Ethiopia,
smallholder farmers in rural areas rely on traditional plants
to preserve the quality of raw cow milk. Such indigenous
knowledge is unevenly distributed among each community
member. Location, religion, linguistic and cultural
backgrounds are among the factors that influence the use of
herbal plants for preserving raw milk (Ashenafi and
Beyene, 1993). The knowledge of medicinal plants is
oral as well as written documentation.
Ethiopia has more than 6500 higher diverse flora and the
country has one of the six plant biodiversity-rich regions
(Egziabher, 1991). Some of these plants have been
traditionally used to preserve raw and fermented milk, for
example, Lippia javanica, Olkingiri, and Olea Africana
used by the Maasai community in Kajiado district to
process and preserve milk were collected from the field
(Onyango et al., 2015). There is considerable potential for
the utilization of natural antimicrobials in food, especially
in fresh milk (Ashenafi and Beyene, 1993; Gaysinsky et al.,
2007). Timely cooling ensures that the quality of the milk
remains good for processing and consumption (Connor,
1995). However, in rural places where there is no
refrigerator facility people use traditional approaches to
maintain the quality and safety of milk. In different parts of
Ethiopia use of herbs and plants as milk preservation or as

26
 27
milk container fumigation is a common practice for a long
period. The use of wood smoke provides a component of a
hurdle system for milk preservation. Among the functional
components of smoke phenols and acids have been the
most antimicrobial activity, while carbonyls and acids have
a wide spectrum of antibacterial activity even at low levels
of phenols (Toledo, 2008; (Wanjala et al., 2016b).
However, there was no documented study on the role
of these herbs and plants and all herbs and plants are
considered equally effective in improving the quality,
safety, and shelf life of milk. So, the objective of this study
is to evaluate the most common herbs and plants
traditionally used in different parts of the country to clean
and fumigate milk equipment to enhance the safety and
quality of milk and milk products.
MATERIALS AND METHODS
Study Area
Holeta agricultural research center, National
Agricultural Biotechnology Research Center under
microbial research laboratory.
Collection and Preparation of Cleaning and Smoking
Herbs and Plants
In this study, the following most common cleaning and
smoking herbs and plants most commonly used in the
central highlands of Ethiopia that was identified previously
by the East African Agricultural Product (EAAPP)
assessment study were used.
Cleansing Herbs
1 Cucumis myriocarpus
2 Thymus shrimperi
3 Zehneria scabra
Smoking Plants
1 Olia Africana
2 Terminaliia brownie fres
3 Olinia rochetiana
These herbs and plants were collected following the
same procedures as traditionally used in the community.
The steam of these plants was chopped in 5 to 10cm for the
smoking of the utensils and the leaf with steam before
drying of these herbs were used for cleaning the inner of
the milk utensil with clean tap water.
Milk Sampling
The milk sample was taken from the Holeta
Agricultural Research Center (HARC) dairy farm of a
healthy udder and free of any clinical mastitis cow. The
milk was filtered with sterile cheesecloth to remove
physical debris like animal dung, feed, and hair residue in
the milk.
Treatment
T1= Cucumis myriocarpus X Olia Africana
T2= Cucumis myriocarpus X Terminaliia brownie fres
T3= Cucumis myriocarpus X Olinia rochetiana
T4= Thymus shrimperi X Olia Africana
T5= Thymus shrimperi X Terminaliia brownie fres
T6= Thymus shrimperi X Olinia rochetiana
T7= Zehneria scabra X Olia Africana
Inter J Appl Sci Engr, 2021, 9(1): 26-30.
T8= Zehneria scabra X Terminaliia brownie fres
T9= Zehneria scabra X Olinia rochetiana
T10= Control
Milk Utensils Used
In this study plastic pots of wider mouth with a cap
having mostly used in the community and easy for cleaning
and fumigation were used. The containers were capable of
holding up to two liters of milk.
Cleaning and Fumigation of Milk Equipment
The inner surface of milk storage equipment was
cleaned effectively with the cleaning herbs separately using
clean tap water. Fumigation of milk equipment was done
with the air-dried smoking plant with sufficient amounts
for 20-30 minutes to produce enough smoke. Fumigation
of the equipment with the plant steam was done separately
following the traditional practice. Equipment cleaned with
only tap water and not fumigated was used as a control.
The raw milk of 400 ml was added to each fumigated
container and the control container and stored at room
temperature.
The Microbial Quality and Growth Dynamics during
the Storage of Raw Milk
In this study, the effect of these treatments on the
microbial growth dynamics was tested using the most
common indicator microbes of a total aerobic mesophilic
bacterial count, coliform count, and lactic acid bacterial
growth test was done at 0 hrs, 8 hrs, and 25 hrs during
storage of milk at room temperature.
Total Aerobic Mesophilic Bacterial Count
A serial dilution of milk samples was done from 101-
10 . From 103 and 104 dilution 1ml was proud to 15x90 mm
4
plate. Next 20ml of sterile plate count agar (PCA) (Oxoid)
medium was poured and mixed with sample by rotating
clockwise and anticlockwise. The plates were allowed to
cool in a laminar flow hood for 15 minutes and incubated
aerobically at 37 °C for 48 h. Finally, the colony counting
was done using a digital colony counter and the counts
were converted into cfu/ml recorded as total aerobic
mesophilic bacterial count.
Total Coliform Counts
From serial dilution of 101 and 102 one milliliters was
poured to a 15x90 mm plate in duplicate and 20ml of sterile
Violet Red Bile Agar (Oxoid) medium was added and mixed
with the sample and allowed to solidify for 15 minutes in a
laminar flow hood. Finally, the plates were inverted and
incubated at 30°C for 24 hrs and typical red colonies on the
plates were considered coliforms and counted.
Mesophilic Lactic Acid Bacterial Count
Isolation of LAB was done according to the standard
method of ISO 15214:1998. In detail, from a serial dilution
of 101 and 102 one milliliters of the sample was poured to
15x90 mm plate in duplicates and 25 ml of MRS (de-Mann,
Rogosa and sharp, Oxoid) The medium was allowed to
solidify for 10 minutes. Finally, the plates were inverted
and incubated anaerobically, using anaerobic jars, at 30 oC
for 48 hours. Colonies growing in a medium were counted
after incubation at 30°C for 72 h.
 28
Physicochemical Parameter
Titratable acidity and pH was tested at 0, 8, and 24 hrs
intervals to estimate the physicochemical stability of the
raw milk stored in these treated milk utensil.
Titratable Acidity
Titratable acidity of each treatment was assayed using
the titration method (ISO & IDF, 2012). Titratable acidity
was the measure of free and bound hydrogen ions by
titrating with 0.1 N NaOH and expressed in percent (m/v).
Each sample was titrated with standard 0.1 N NaOH using
4 drops of 0.1% phenolphthalein as color indicator until
faint color persists, the end-point has been reached at pH of
8.2 (Fabro et al., 2006).
Titratable acidity produced during fermentation was
calculated as:
Titratable acidity (TA)%=(V NaOH in liter x 0.1N x
90.08 g/mol)/(10 ml) x 100%
Where: V is the volume of NaOH consumed, 0.1 N is the
concentration of NaOH in normality, 90.08 g/mol is the
molecular weight of lactic acid.
pH Test
The pH of each treatment was measured according to
the standard methods of ISO 26323:2009(E). The pH of the
samples was measured by the potentiometric method i.e. by
a potential difference between the sample and electrolyte
solution present inside the electrode of the pH meter using
a digital pH meter. First, the pH meter was calibrated with
fresh pH 4.0 and 7.0 standard buffer solution (Tomovska et
al., 2016). The electrode of the pH meter will be directly
dipped into the milk sample of treatment and the pH
reading was recorded.
Sensory Preference Test
For consumer preference, pasteurized milk of 400 ml
was added to equipment treated with the cleansing herbs
and smoking plants separately and milk in a control.
Pasteurized milk stored in each container cleaned and
fumigated with each herb, the plant was provided for
consumers, and their preference was scored concerning the
control. Each treatment sample was randomized and coded
with three-digit numbers and served in plastic cups
separately following the methods of (Yang and Li, 2010).
A test form comprising five sensory attributes was given to
each assessor. The sensory assessments were conducted by
involving ten semi-trained panelists of adult males and
females. The panelists were provided with a glass of water
and, instructed to rinse with water between while working
with the samples. A written instruction was given to a
panelist to evaluate the products for odor, test, and overall
acceptability using a five-point hedonic scale: 1 = Dislike
Extremely, 2= Dislike Moderately, 3= neither like nor
dislike 4= Like Moderately, and 5= Like Extremely. In the
evaluation of the sensory characteristics of the product, one
was equal to the worst and five was equal to the best.
Experimental Design
CRD with a factorial in triplicate in which Three types
of cleaning plant and three types of smoking plant were
used in this experiment.
Inter J Appl Sci Engr, 2021, 9(1): 26-30.
Statistical Analysis
Data obtained from the laboratory test were analyzed
using Statically Analysis Software (SAS) ver 9.4; 2014 and
Minitab 20.3 ver. 2020. Analysis of variance means
separation within and between treatment was done using
Tukey's test at α=0.05 significance level after confirmation
of the normality of data.
RESULTS AND DISCUSSION
Titratable acidity and pH analysis results of raw milk
stored at room temperature were summarized in Table 1.
The summary table showed, the efficacy of different milk
equipment cleaning herbs and smoking plants in
maintaining the change in physicochemical properties, pH,
and titratable acidity of raw milk stored at room
temperature. The result indicated titratable acidity and pH
were varied during storage. However, treatment T1, T4,
and T7 were no significant (P>0.05) change in pH and
treatable acidity during 0, 8, and 24 hrs of storage at room
temperature(25±3oC).
The pH and treatable acidity of treatment T8, T9, and
control samples were a significant change during 0, 8, and
24 hrs of storage. The titratable acidity of treatment T6 was
not significantly changed during the first 8 hrs but later
during the 24 hrs of storage, they were significantly (p <
0.05) increased.
The microbial growth of total mesophilic bacteria,
coliform, and mesophilic lactic acid bacteria (LAB) in milk
stored in equipment treated with T1, T4, and T7 had no
significant (α=0.05) growth change during storage of the
raw milk at room temperature Table 2. On the other side
total mesophilic bacteria, coliform, and lactic acid bacteria
in treatment T2, T6, T9, and control samples were
significant growth change during 8 and 24 hrs of storage at
room temperature. The microbial growth of mesophilic
bacteria count had 0.05-0.07 log unit change in T1, T4, and
T7 respectively during 24 hrs of storage.
The results of the control sample reinforce the general
belief that the increase of microbial population in food
during storage of food product at ambient temperature.
Total plate count, total coliform, and lactic acid bacteria
maximum growth of 0.90, 1.02, and 1.12 log units were
observed during 24 hrs of storage in a control sample.
Table 3 showed the sensory acceptability of milk
stored in utensils treated with different cleaning herbs and
smoking plants. All treatments were significantly better
color acceptance than the control milk sample stored in
unsmoked utensils. Raw milk stored in milk equipment of
treatment T1 was significantly better odor acceptance than
treatment T2, T5, T8, T9, and the control sample. Smoking
was capable of changing the odor and color of milk. The
overall sensory acceptance of the milk stored in milk
utensils treated with T1, T4, and T7 had relatively better
consumer acceptance Table 3.
Titratable acidity and pH were the common indicators
of the keeping quality(freshness) of raw milk (Schmidt and
Shirley, 2014). It helps to estimate the physical, chemical,
and microbiological stability of the food(Tomovska et al.,
2016). The pH of raw milk ranged from 6.64-6.68 and
titratable acidity was 0.14 to 0.16%(Connor, 1995; Tesfay,
2015). However, during the storage of milk, the pH was
decreased because of the fermentation of lactose to lactic
 29 Inter J Appl Sci Engr, 2021, 9(1): 26-30.

Table 1: Change in titratable acidity and pH of milk during storage at room temperature
pH TTA
Treatment 0hrs Mean±SD 8hrs Mean±SD 24hrs Mean±SD 0hrs Mean±SD 8hrs Mean±SD 24hrs Mean±SD
T1 6.67±0.03a 6.58±0.02a 6.59±0.02a 0.14±0.01a 0.14±0.02a 0.15±0.02a
a ab b c b
T2 6.67±0.02 6. 26±0.05 5.59±0.03 0.14±0.01 0.25±0.01 0.31±0.01a
T3 6.67±0.01a 6.30±0.03ab 5.84±0.3b 0.14±0.01c 0.25±0.01b 0.31±0.01a
a a a a a
T4 6.67±0.03 6.67±0.06 6.66±0.03 0.14±0.02 0.15±0.00 0.16±0.01a
T5 6.67±0.04a 6.34±0.14a 5.81±0.01b 0.14±0.01c 0.28±0.01b 0.31±0.01a
T6 6.67±0.02a 6.27±0.01a 5.86±0. 2b 0.14±0.01b 0.16±0.01b 0.29±0.01a
T7 6.67±0.03a 6.66±0.2a 6.66±0.06a 0.14±0.02a 0.15±0.02a 0.15±0.01a
T8 6.67±0.01a 6.37±0.07b 6.04±0.04c 0.14±0.01c 0.24±0.01b 0.30±0.02a
a b c c b
T9 6.67±0.05 6.22±0.02 5.95±0.11 0.14±0.01 0.25±0.03 0.32±0.02a
Cont 6.67±0.03a 5.93±0.07b 4.76±0.12c 0.14±0.01c 0.32±0.02b 0.39±0.01a
Where T1, T2…T9= treatment; TTA= Total titratable acidity; control = without any treatment, means across a raw that do not share a
letter are significantly different.

Table 2: Efficacy of each treatment on stabilizing the growth of microorganisms in milk

Trt PCA (log10CFU/ml) Mean ± SD Coliform count (log10CFU/ml) Mean ± SD LAB (log10CFU/ml) Mean ± SD
0hr 8hr 24hr 0hr 8hr 24hr 0hr 8hr 24hr
T1 5.12±0.03a 5.17±0.05a 5.19±0.01a 3.63±0.01a 3.64±0.02a 3.66±0.02a 3.11±0.09a 3.17±0.05a 3.23±0.02a
T2 5.14±0.04c 5.31±0.05b 5.44±0.01a 3.37±0.01c 3.84±0.09b 4.13±0.06a 3.36±0.01c 3.92±0.03b 4.25±0.04a
a b b c b a
T3 5.06±0.01 5.18±0.03 5.20±0.53 3.56±0.02 3.91±0.03 4.16±0.06 3.58±0.01c 3.91±0.06b 4.13±0.11a
T4 5.24±0.01a 5.25±0.03a 5.29±0.01a 3.80±0.01a 3.81±0.01a 3.82±0.01a 3.92±0.01a 3.92±0.02a 3.95±0.01a
b ab a b b a
T5 5.28±0.01 5.34±0.01 5.40±0.04 3.83±0.01 4.02±0.08 4.69±0.13 3.59±0.01c 3.81±0.06b 4.26±0.06a
T6 5.58±0.03c 3.59±0.01b 3.74±0.03a 3.86±0.00c 4.03±0.08b 4.62±0.06a 3.60±0.01c 3.98±0.01b 4.39±0.12a
T7 5.36±0.01a 5.38±0.01a 5.41±0.05a 3.93±0.01a 3.93±0.01a 3.94±0.01a 3.65±0.01a 3.66±0.01a 3.67±0.01a
b b a c b a
T8 5.06±0.01 5.20±0.05 5.45±0.12 3.90±0.01 4.29±0.19 4.88±0.09 3.59±0.01c 3.86±0.07b 4.15±0.06a
T9 5.05±0.01c 5.32±0.01b 5.43±0.01a 3.92±0.01c 4.34±0.09b 4.92±0.04a 3.58±0.01c 3.95±0.05b 4.30±0.07a
c b a c b a
Cont 5.01±0.01 5.52±0.03 5.91±0.02 3.81±0.02 3.92±0.03 4.83±0.02 3.62±0.01c 4.22±0.12b 4.70±0.01a
Where T1, T2…T9= treatment; TTA= Total titratable acidity; control = without any treatment; 0hr= initial hours, and Means across a
raw that share a letter are not significantly different.

Table 3: Effect of treatment on the organoleptic properties of raw milk in each treatment
Treatment Odor (Mean ±SD) Color (Mean ±SD) Test (Mean ±SD) Overall (Mean ±SD)
T1 4.10±0.01ab 4.47±0.06a 4.17±0.21a 4.24±0.11ab
ab bc a
T2 3.93±0.15 3.83±0.35 3.87±0.32 3.88±0.11d
T3 3.80±0.10b 3.93±0.20ab 3.73±0.12ab 3.82±0.07d
a ab a
T4 4.30±0.10 4.13±0.15 4.23±0.15 4.22±0.05abc
T5 4.03±0.21ab 3.70±0.10bc 4.00±0.20a 3.91±0.10cd
T6 3.90±0.14ab 3.85±0.21abc 3.95±0.07a 3.90±0.14bcd
a ab a
T7 4.33±0.15 4.23±0.06 4.23±0.25 4.27±0.12a
T8 3.97±0.15ab 3.87±0.11b 3.83±0.15ab 3.89±0.13d
ab bc a
T9 3.97±0.15 3.80±0.26 4.07±0.12 3.94±0.16bcd
Control 3.13±0.06c 3.30±0.20c 3.30±0.10b 3.24±0.08e
Where T1, T2…T9= treatment, SD= standard deviation; column sharing the same letter was no significant difference

acid by lactose fermenting microorganisms(Schmidt and
Shirley, 2014). In addition, as the storage time increased
lactic acid and other organic acids were accumulated and
the total titratable acidity of the products was increased. As
the pH falls and total titratable acidity were raised, the
whey protein begins to coagulate and forms a curd. So,
preventing or stabilizing the growth of lactose fermenting
microbes in milk helps to prevent coagulation and improve
the shelf life or the keeping quality of raw milk.
Accordingly, treatment T1, T4, and T7 were no
significant change in pH and titrable acidity at initial, 8, and
24 hrs of storage at a room temperature of 25±3 oC storage.
This might be due to the treatment effect of the growth of
these microbes. The cleaning and smoking plant stabilizes
the growth of the microbes rather than inhibiting the
growth. Related results of insignificant change in pH and
treatable acidity during 24 hr storage of raw milk stored in
cleaned and smoked equipment were reported by (Ashenafi
and Beyene, 1993). (Bekuma et al., 2018), also reported
that milk stored in milk utensils smoked with different
plants were low acid production during storage. This
implies that these treatments used in this study were better
efficacy of preserving raw milk in areas having no cooling
facilities, no facilities of transportation, and no market
access. Concerning the sensory acceptability of the
products, and overall acceptability of milk stored in treated
milk utensils was better than the control sample. This might
be due to the aroma of the smoke diffusing into the milk
and improving the flavor of milk. Bekuma et al., (2018)
reported that smoking milk utensils with different plants
improve the aroma and shelflife of milk.
The use of smoking plants and cleaning herbs for milk
equipment hinder or stabilize the growth of microflora in
milk. In this study, there is no significant growth of
microbial in milk stored in milk equipment stored in
treatment T1, T4, and T7. This might be due to the
antimicrobial effect in the smoke of these plants and
cleaning herbs. Related results were reported by (Wanjala
et al., 2016a).
Conclusion
This study evaluates the traditional methods used to
preserve and improve the organoleptic properties of cow
raw milk in Ethiopia. Accordingly, the most common milk
 30
storage equipment cleaning herbs and smoking plants used
in different parts of Ethiopia were evaluated for their
preservative role. The efficacy of these treatments in terms
of stabilizing the growth of microorganisms, changes in
physicochemical properties like pH, treatable acidity, and
organoleptic improvement capability. Among these nine
treatment treatments T1, T4 and T7 were relatively better
at keeping the quality of raw milk. So, utilization of these
treatments was better to improve the keeping quality of raw
milk especially for household and small-scale milk
producers of the rural area where there is no cooling facility
and limitation of transport to sell or deliver raw milk to
shopping on time. In addition, utilization of these
treatments has a role in reducing postharvest loss due to
microbial spoilage.
Acknowledgments
Ethiopian Institute of Agricultural research center
Food Science and Nutrition Research Directorate for
financial as well as reagent support. Holeta Agricultural
Research center, dairy research Laboratory for lab
facilities.
Authors Contribution
Zerihun Asefa has developed project concept, did
overall research plan, data collection, statistical analysis;
report writing & primary responsibility for final content.
Esayas Abrha was developing the overall research plan,
supervision, providing essential reagents, and editing the
final manuscript.
Conflict of Interest
All the authors confirm that there is no conflict of
interest.
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